Reduced-calorie fats containing behenic acid and process for preparing same

ABSTRACT

The invention relates to a reduced calorie fat and a process for the preparation of a reduced calorie fat, which includes esterification of edible oils with a source of behenic acid in the presence of a catalyst at a temperature in the range of about 25° C. to 150° C. for at least about 0.5 hours, and then recovering and purifying the reduced calorie fat using conventional methods.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application is a divisional of U.S. Ser. No. 09/207,056, nowU.S. Pat. No. 6,617,141, the entire content of which is incorporated byreference, and which patent claims priority to Indian Application No.3504/DEL/97, filed Dec. 8, 1997.

FIELD OF THE INVENTION

[0002] This invention relates to a process for the preparation ofreduced calorie fats. This invention particularly relates to thepreparation of reduced calorie fats by incorporating behenic acid intoedible oils such as sunflower, groundnut, safflower, rapeseed, soybeanand fish oils. More particularly, it provides a reduced calorie plasticfor containing essential fatty acids and natural antioxidants usingdifferent processes involving chemical interesterification of1,3-dibehenin and edible oils or chemical interesterification of1,3-dibehenin based structured fat and edible oils or enzymictransesterification of edible oils with alkyl behenates.

BACKGROUND OF THE INVENTION

[0003] Typical fats and oils provide approximately 6 kcal/g ofmetabolizable energy compared to 4 kcal/g for protein or carbohydrate[Atwater, W. O. et al., Annu. Rep. Storrs Agric. Exp. Stn. (1903) No.15,123-146 & Maynard, L. A., J. Nutr. 28 (1944) 443-453]. In addition tothe caloric and nutritional value, fats have many functions in the diet.Fats and oils carry, enhance and release the flavours of other foodcomponents, delays digestion, increases palatability of food and impartsthe feeling of satiety. Certain unsaturated fatty acids like9,12-octadecenoic acid (linole acid) which are known to be essentialfatty acids are necessary as they are not produced in the body. Fats andoils are also associated with the fat-soluble vitamins A, D, E and K,and the absorption of these vitamins is impaired at very low fatintakes. Fat is also associated with diseases such as coronary heartdisease and cancer, a high fat diet being positively linked to both. TheU.S. Surgeon General has recommended that no more then 30% of thedietary calories should be derived from fat [U.S. Department of Healthand Human Services, The Surgeon General's Report on Nutrition andHealth, DHHS (PHS) Publication 88-50210, [U.S.GPO., Washington, D.C.(1988)]. Regulatory and advisory bodies advocate a lowered fat intake inorder to reduce the incidence and morbidity of many coronary diseases,stroke, high blood pressure, obesity and diabetes.

[0004] However, food habits are difficult to change and the positivecontribution of fats to increase the palatability of foods is generallyrecognized. The level of fat in the diet of affluent societies is toohigh and needs to be lowered. Reduced fat or low calorie foods as wellas fat replacers or substitutes have been the result of numerousattempts to meet the health recommendations without changing traditionalways of eating.

[0005] Three different types of fat replacers namely carbohydrate-based,protein-based and fat-based are reported in the literature.

[0006] Carbohydrate-based fat replacers consists of many products likedextrin, polydextrose, maltodextrin, cellulose, gums etc., which areused as thickeners and stabilizers in frozen desserts, salad dressings,margarine type spreads, baked products, frostings and snacks. Manyproducts based on starch have been developed specifically as fat mimetic[Alexander, R. J., Cereal Food World 40 (1995) 366-368]. However, starchbased products are not good for diabetics for whom good glucose controlis necessary [Grundy, S. M., Diabetics Care 14 (1991) 796-801]. The U.S.Food and Drug Administration (FDA) regulations state that sensitiveindividuals may experience a laxative effect from excessive consumptionof maltrodextrins [U.S.A. Food & Drug Administration Regulations 21 CFRPart 105]. Some popular examples of carbohydrate based fat replacers areStellar, Remyrise AP, N-Oil, Lycadex, Maltrin, Ex-cel, Fibercel, CentuTex, Fibrex etc. [Jones, J. M., Chemistry & Industry, (1996) 494-498].

[0007] Protein-based fat replacers are produced using common proteinssuch as egg white, skimmed milk or whey by microparticulating them intoa particle size of 1-3 μm to obtain a slippery and creamy fat likefeeling which provide 1-2 kcal/g [Singer, N. S., et al., J.] Amer. Coll.Nutr. 9 (1990) 388-397]. These products are being used in variety offood products such as yogurts, cheese products, frozen desserts and alsofor formulating low fat baked goods such as cheese cakes and pie crust.Gelatin from fish waste was also reported as a fat replacer to use as aviscosity modifier and to impart a creamy texture. However, such fatreplacers are reported to be hypersensitive for persons with allergy tothe base proteins [Young, V. R., et al., J. Amer. Coll. Nutr. 9 (1990)418-426]. Some examples of protein based fat replacers popular in themarket are Dairylight, Simplesse, Lita, Calpra 75 etc. [Jones, J. M.,Chemistry & Industry (1996) 494-498].

[0008] Carbohydrate-based and protein-based fat replacers are presentlyused in a range of foods, and are effective in delivering fat-liketexture where the final product has a significant water content and isnot exposed to extremely high temperatures or temperature variations[Mela, D. J., Fett/Lipid 98 (1996) 50-55]. These problems can beovercome by fat-based low calorie fats.

[0009] Fat-based low calorie fats have many advantages like functionaland sensory properties very similar to the normal fats when compared tothe carbohydrate and protein-based fat replacers. They also provide boththe characteristic texture and flavour effects of native fats.

[0010] Many fat-based low calorie fats are reported in the literaturenamely propoxylated glycerols esterified with fatty acid chlorides[Masten, L. W., EP 571,219 (1993); White, J. F. et al., EP 325,010(1989)]; fatty acid diesters of C₄₋₁₀ dihydric alcohols [Klemann, L. P.et al., U.S. Pat. No. 5,286,512 (1994); Klemann, L. P. et al., U.S. Pat.No. 5,006,351 (1991)]; trioltriester derivatives [Klemann, L. P. et al.,U.S. Pat. No. 5,043,179 (1991)]; polyol fatty acid polyesters [Kester,J. J. et al., U.S. Pat. No. 5,314,707 (1994); Letton, J. C., et al.,5,306,514 (1994)]; polyvinyloleate [D' Amelia, R. P. et al., U.S. Pat.No. 4,915,974 (1990)]; oleoylloeate [Jacklin, P. T. et al., U.S. Pat.No. 4,915,974 (1990)]; bis-oleoylaspartyladipare [Klemann, L. P. et al.,U.S. Pat. No. 5,139,807 (1992)]; esterified alkoxylated mono- anddiglycerides [Cooper, C. F. et al., U.S. Pat. No. 5,371,253 (1994)];triglycerides containing C₁₂₋₂₂ fatty acids having alkyl groups at leastat the position 5,9,13 of the alkyl chain [Tagiri, M. et al., JP04,325,055 (1992)]; 1,3-didecanoylglycerol [Mazur, A. W. et al., U.S.Pat. No. 5,137,660 (1992)]; alkyl or polyol thioesters [Klemann, L. P.,U.S. Pat. No. 4,992,293 (1991)]; propyleneglycol diesters of mediumchain and long chain saturated fatty acids [Stipp, G. K. et al., EP495,553 (1992)]; alkylmalonic acid diesters [Fulcher, J. G. et al., Aus.Pat. 594,040 (1990)]; esterified polyoxyalkylene block co-polymers[Cooper, C. F. et al., EP 481,717 (1992)]; alkylglycoside fatty acidpolyesters [Winter, D. D. et al., U.S. Pat. No. 4,942,054 (1990)]; fattyacid esters of sucrose [Letton, J. A., et al., EP 375,027 (1990)];sorbitol fatty acid esters [Gruetzmacher, G. D., EP, 591,258 (1994)];partially esterified polysaccharide with fatty acids [White, J. F. U.S.Pat. No. 4,959,466 (1990)]; alkoxylated sugar and sugar alcohol esters[Ennis, J. L. et al., EP 425,635 (1991)]; polysaccharide fatty acidpolyester [Meyer, R. S. et al., U.S. Pat. No. 4,973,489 (1990)]. Allthese are unnatural compounds not normally encountered in human diet andthe long term affects of consumption of such are presently unknown.

[0011] Examples of the more commonly known low calorie fats areOLESTRA™, medium chain triglycerides (MCTs), Caprenin and SALATRIM™.OLESTRA™ is a mixture of hexa-, hepta-, and octa-fatty acid esters ofsucrose. The physical properties of sucrose polyesters are similar tonormal triglycerides [Jandacek, R. J., et al., Chem, Phys. Lipids 22(1978) 163-176]. OLESTRA™ is adaptable to most application where fatsand oils are used. However, the major drawback to OLESTRA™is “analleakage”, the result of a non-digestible fat passing through thedigestive system. OLESTRA™ also blocks the absorption of fat solublevitamins [Bailey's Industrial Oil & fat Products, Vol. 1, Ed. Y. H. Hui(1996), p. 286; Jones D. Y. et al., Amer. J. Clin. Nutr. 53 (1991)1282-1287 and Dasher, G., et al., FASEB J. 8 (1994) 443].

[0012] MCTs are triglycerols composed of C₆, C₈ and C₁₀ saturated fattyacids. Hunder, J. E., et al., [U.S. Pat. No. 4,863,753 (1989)] reporteda low calorie peanut butter containing ≧10% MCTs. A peanut buttercomposition containing MCTs (41.96%) was reported to have excellentconsistency and contained at least 10% fewer calories than the normalfat. However, MCTs may be toxic and may induce metabolic acidosis inlarge doses [Akoh, C. C. Inform 6 (1995) 1055-1061].

[0013] SALATRIM™ (short and long acylglycerol molecules) is a family ofstructured triacylglycerols prepared by interesterifying a completelyhydrogenated vegetable oil with triacetin, tripropionin and/ortributyrin using sodium methoxide as a catalyst at 100-150

C [Wheeler, E. L. et al., U.S. Pat. No. 5,258,197 (1991); Klemann, L. P.et al., 42, J. Agr. Food Chem. (1994) 42, 442-446]. Thus, the SALATRIM™triglycerides are composed of mixtures of long-chain saturated fattyacids (predominantly stearic) and short-chain fatty acids (aceticpropionic, and/or butyric) esterified to the glycerol backbone. Asimilar low calorie fat namely acetyl distearoyl glyceride was preparedby Wheeler, E. L. et al. [U.S. Pat. No. 5,258,197 (1991)]. Extensivetesting in animals of SALATRIM™ has shown no changes in the intestinalmicroflora or secondary bile acids, and no increased mutagenicity orother toxicological effects [Hayes, J. R., et al., J. Agr. Food Chem. 42(1994) 500-514; Scheinbach, S., et al., ibid, 42 (1994) 572-580; Hayes,J. R., et al., ibid, 42 (1994) 539-551; Hayes, J. R., et al., ibid, 42(1994) 515-520; and Hayes, J. R., et al., ibid, 42 (1994) 521-527].However, SALATRIM™ contains unnatural components such as very lowmolecular weight fatty acids and does not contain essential fatty acids.

[0014] Caprenin, a structured triglyceride consisting of caprylic andcapric acids and the very long chain behenic acid which was developedfor use in chocolate preparation. It yields only 5 cal/g instead of 9cal/g because the short chain fatty acids have lower energy values andbehenic acid is not well absorbed [Peters, J. C. et al., J. Am. Coll.Toxicol. 10 (1991) 357-67; Webb, D. R., et al, ibid 10 (1991) 341-356;Webb, D. R. et al., ibid., 10 (1991) 325-340]. Glycerin was esterifiedfirst with behenic acid to form glycerylmonobehenate, which was thenreacted with capric and caprylic acids or their anhydrides, and thenpurified by molecular distillation and steam deodorization. Yoshida, T.et al., [JP 0559392 (1991)] reported the synthesis of2-behenyl-1,3-dicaproyl glycerol by reacting tribehenin and ethylcaproate in the presence of lipase and the product's feed study on ratsshowed that absorption of the structured triglycerides was significantlylower and its excretion into feces was higher than other oils. Inanother report Yoshida R., et al., [Shoka to Kyushu 14 (1991) 27-30 C.A.117: 47249 (1992)] reported the synthesis of triglycerides with arandomly placed long chain fatty acid (behenic acid) and two mediumchain fatty acids (capric and caproic acids) and included it in the dietof rats. These triglycerides were poorly absorbed from the intestine;absorption of behenic acid was particularly poor. Caprenin feedingstudies have shown that it produces no toxic effects when fed as theprimary source of dietary fat [Webb, D. R. et al., Food Chem. Toxicol.31 (1993) 935-946]; Webb, D. R., et al., J.Amer. Coll. Toxicol. 10(1991) 341-356 and Webb, D. R., et al., ibid, 10 (1991) 325-340].However, a six week study of relative effects on serum lipids andapolipoproteins of a caprenin rich diet feeding studies by Wardlaw, G.M., et al., [Am.J. Clin. Nutr. 61 (1995) 535-542] showed that caprenincan contribute to hypercholesterolemia in men and gastrointestinalcomplaints in some individuals.

BRIEF DESCRIPTION OF THE DRAWINGS

[0015]FIG. 1A shows Table 1A, which is a graphical representation ofmean body weight gain of all the test groups including the ad libitumgroup in the restricted diet growth experiment.

[0016]FIG. 1B shows Table 2B, which is a graphical representation of thegrowth pattern of the 10% SO (control group) and 10% SL 3 (experimentalgroup) in the ad libitum experiment.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] The low calorie fats reported so far does not contain essentialfatty acids and the natural antioxidants normally found in natural oilsand fats. Hence, the objective of the present invention is to providereduced-calorie fats which fulfils the three basic functions of fat infoods: (1) a source of essential fatty acids, (2) a carrier for fatsoluble vitamins, and (3) a source of energy for storage or oxidation.This is achieved by incorporating poorly absorbable behenic acid intoedible oils particularly vegetable oils such as sunflower, safflower andgroundnut oils. A further objective of the present invention was toprovide a reduced calorie plastic fat of the consistency of vanaspatiand which does not contain the deleterious trans fatty acids.

[0018] In the present invention, reduced-calorie fats were prepared byincorporating behenic acid into edible oils using three differentroutes.

[0019] In the first two methods for the preparation of reduced-calorieplastic fats, 1,3-dibehenin prepared from mustard oil was used as asource of behenic acid. Other cruciferae oils such as rapeseed oil couldalso be used for this preparation. Mustard oil consists of about 44% oferucic acid along with other normal fatty acids. 1,3-Dierucin isprepared from mustard oil using the methodology developed by uspreviously [Kaimal T. N. B., et al. Biotechnology Letters 15, (1993)353-356)] by lipase (from Candida cylindracea) hydrolysis underrestricted water conditions and isolated 1,3-dierucin from the reactionmixture by crystallization from acetone at 10° C. 1,3-Dierucin ishydrogenated to 1,3-dibehenin using conventional method with 10%palladium-carbon as catalyst under pressure (2-3 kg/cm²) in chloroform.After removal of the catalyst by filtration, 1,3-dibehenin isrecrystallized from acetone.

[0020] The first route comprises interesterification of vegetable oils,exemplified by sunflower oil, and 1,3-dibehenin using sodium methoxideas catalyst at a concentration in the range of 0.3% to 0.5% by weight ofthe substrate at a temperature in the range of 80-150° C. for a periodin the range of 0.5 to 1.0 hour, purifying the product by silicic acidcolumn chromatography to yield a reduced-calorie fat. The product thusprepared from sunflower oil and dibehenin in the molar ratio of 1:0.5contained 29.8% of behenic acid and 44.6% of linoleic acid while aproduct containing 52.5% of behenic acid and 24.8% linoleic acid wasobtained when the molar ratios were 1:1.

[0021] The second route provides a process for the preparation ofreduced-calorie fat which comprises incorporation of sunflower oil fattyacids into the second position of 1,3-dibehenin to prepare a structuredfat followed by its chemical interesterification with vegetable oilsexemplified by sunflower oil. Accordingly this route consists of thefollowing steps: a) saponification of sunflower oil to obtain fatty acidmixture, b) conversion of fatty acid mixture to anhydrides c)esterification of 1,3-dibehenin with the said fatty acid anhydrides toobtain structured fat, d) chemical interesterification of the structuredfat with vegetable oils exemplified by sunflower oil, and e) isolationof reduced-calorie fat by silicic acid column chromatography.

[0022] Sunflower oil is saponified using conventional method byrefluxing with 10% potassium hydroxide in ethanol and the saponifiedmass was neutralised with dilute hydrochloric acid and extracted withdiethylether to obtain sunflower oil fatty acids. The sunflower oilfatty acid mixture was converted to their anhydride by treating withdicyclohexylcarbodiimide following conventional procedures. The fattyanhydride was then refluxed in chloroform with 1,3-dibehenin in presenceof N,N-dimethylamino pyridine to obtain structured fat. The structuredfat with a melting point of 55° C. was then interesterified withvegetable oils exemplified with sunflower oil using sodium methoxide ascatalyst at a concentration in the range of 0.3% to 0.5% by weight ofthe substrate and at a temperature in the range of 80-150° C. for aperiod in the range of 0.5 to 1 hr till the randomization of the fattyacids was complete. The reduced-calorie fat was then purified by silicicacid column chromatography and found to contain about 32.1% of behenicacid and 33.7% of linoleic acid with a melting point of 36° C.

[0023] The third route of the present invention provides a process forthe preparation of reduced-calorie fat which comprisestransesterification of vegetable oils and alkyl behenate with lipase[from Mucor miehei (Lypozyme 1M 20), 30 BIU/g, supplied by Novo IndustriA/S, Denmark] and purifying the structured fat by silicic acid columnchromatography to yield a reduced-calorie structured fat containing 5 to41% of behenic acid and 33 to 60% of linoleic acid which is an essentialfatty acid. Only a slight excess of alkyl behenate (1.2 mole equivalentsto that of vegetable oil) is used for transesterification. Theinteresterified reduced-calorie fat containing about 33% of behenic acidwas found to have a slip melting point of about 37° C. Various alkylesters of behenic acid tried such as methyl, ethyl, isopropyl andn-butyl were used for this reaction but satisfactory levels ofincorporation of behenic acid was obtained only when the ethyl esterswere used.

[0024] The transesterification of sunflower oil and ethyl behenate isstandardised by varying the enzyme concentration in the range of 1 to 4%by weight of the substrate, temperature in the range of 55-70° C. andreaction period in the range of 1-6 hrs. Of the various parametersstudied, enzyme concentration of 2%, temperature of 60° C. and reactionperiod of 3 hr yielded a structured fat containing about 34% of behenicacid by GC analysis, and having a slip melting point of 37° C.

[0025] Accordingly the present invention provides a process for thepreparation of reduced-calorie fat which comprises esterification ofedible oils with a source of behenic acid such as herein described, inpresence of a catalyst at a temperature in the range 25° C. to 150° C.at least for 0.5 hr. and then recovering and purifying thereduced-calorie fats using conventional methods such as hereindescribed. Edible oils which may be used are such as sunflower,groundnut, safflower, soybean, rapeseed, fish oil etc. The sources ofbehenic acid which may be used are 1,3-dibehenin, alkyl behenate and astructured fat containing behenic acid described herein. Theesterification temperature may be in the range of 25° C. to 150° C.depending on the type of catalyst used which may be an alkali metalalkoxide or a thernostable lipase enzyme such as Lipozyme. Theesterification reaction may be effected in the time range of 0.5 hr to 6hr. Recovery of the reduced-calorie fat may be effected by filtrationfollowed by column chromatographic purification in case of enzymatictransesterification or by washing-off the catalyst alkali metal alkoxidein case of chemical transesterification.

[0026] The following examples illustrate the invention and should not beconstrued as the limit of the invention and the manner in which it iscarried out.

EXAMPLES Example 1

[0027] a) Preparation of 1.3-dibehenin

[0028] Mustard oil (100 gm) was mixed with tert-butanol (50 ml) andlipase from Candida cylindracea (700 mg) was added in small portionswhile stirring continuously. After the enzyme was dispersed uniformly,water was added intermittently at a rate of 3 ml/hr by a peristalticpump for 2 h and stirred afterwards for an additional 12 hours. Themixture was centrifuged to separate the lipase. The reaction product hadan acid value of 55.0. The solvent was removed under vacuum and theproduct dissolved in 300 ml of acetone and crystallized at 10

C to yield dierucin (29 gm; colorless powder, m. pt. 45

C). 1,3-dierucin was then hydrogenated using 10% Pd-C (1.5 gm) inchloroform (100 ml) at 2 kg/cm2 hydrogen pressure for 4 hours. Catalystwas filtered off and 1,3-dibehenin was crystallized by adding acetone(50 ml) and then cooling at 0

C. Crystallized 1,3-dibehenin (24 gm; white flakes; m. pt. 88

C) shows 92% behenic acid by Gas Chromatographic (GC) analysis.

[0029] b) Interesterification of Dibehenin and Sunflower Oil

[0030] Sunflower oil (1 g) was mixed with 1,3-dibehenin (0.421 g) andkept under vacuum at 110° C. for 30 minutes. Then sodium methoxide inmethanol (0.5% w/w; 45 μl) was added and stirred for 1 hour undervacuum. The product was taken in 20 ml of diethylether and washed withwater to remove sodium methoxide. The reduced calorie fat containingbehenic acid was purified by silicic acid column chromatography usinghexane:ethyl acetate (95:5, v/v) as eluant and found to contain behenicacid, 29.8%; oleic acid, 16.7%; linoleic acid, 44.6% and others, 8.9%.The product had slip melting point of 34° C.

Example 2

[0031] Sunflower oil (1 g) was mixed with 110° C. 1,3-dibehenin (0.842g) in a two-necked 25 ml of R.B. flask and kept under vacuum at 110° C.for 30 minutes. Then sodium methoxide in methanol (0.5% w/w; 55 μl.) wasadded and stirred for 1 hour under vacuum. The product was taken in 20ml of diethylether and washed with water to remove sodium methoxide. Thereduced calorie fat containing behenic acid was purified by silicic acidcolumn chromatography using hexane:ethyl acetate (95:5, v/v) as eluantand found to contain behenic acid, 52.5%; oleic acid, 13.1%; linoleicacid, 24.8% and others, 9.6%. The yield of the reduced-calorie fat was1.0 g and its slip melting point was found to be 50° C.

Example 3

[0032] Sunflower oil (5 g) was saponified by refluxing with 10%potassium hydroxide in ethanol (20 ml) for one hour. The saponified masswas neutralised with dil. hydrochloric acid and extracted withdiethylether to obtain sunflower oil fatty acids (4.5 g). The fatty acidmixture (4.5 g) was converted to anhydride by treating it withdicyclohexylcarbodiimide (4.24 g) in carbon tetrachloride (30 ml) forovernight. The precipitated dicyclohexyl urea was filtered out andfiltrate was concentrated to obtain fatty acid anhydride (4.1 g). Theanhydride (4.1 g) was then refluxed with 1,3-dibehenin (5 g) in 50 mlchloroform for 10 hours in presence of N,N-dimethylaminopyridine (1.2g). The resulting structured fat having behenic acid in 1,3 position andsunflower oil fatty acids in 2 position was purified by silicic acidcolumn chromatography by eluting with hexane:ethyl acetate (95:5; v/v).An yield of 6 g of structured fat (melting point 55° C.) was obtainedwith fatty acid composition of behenic, 72.2%; oleic, 8.1%; linoleic,12.5%; others 7.2% by GC analysis. This was further interesterified withsunflower oil. Sunflower oil (5.2 g) was mixed with the structured fat(6 g) and kept the contents under vacuum at 110° C. for 30 min. Sodiummethoxide (0.5% w/w; 0.35 ml) was added and the mixture stirred for onehour under vacuum. The product was taken in diethyl ether (150 ml) andwashed with water to remove the excess sodium methoxide. The resultingreduced-calorie fat (10 g) was found to contain behenic acid, 32.1%;oleic acid, 22.1%, linoleic acid, 33.7% and others, 12.5% with a slipmelting point of 36° C.

Example 4

[0033] Sunflower oil (2.5 g) and 1.22 g of ethyl behenate were taken ina screw-capped test tube and the mixture was homogenized by agitationfor 15 min. at 60° C. About 75 mg (2% by wt. of the substrate) ofLipozyme 1M 20 was added followed by agitation of the mixture for 6 hrat 60° C. After the reaction, the enzyme was filtered out and the enzymewas washed with hexane for its re-use. The product was purified bysilicic acid (60-120 mesh) column chromatography. The ethyl esters wereeluted by using hexane:ethyl acetate (99:1, v/v) followed bytriglycerides using a solvent mixture of hexane:ethyl acetate (95:5,v/v). The triglycerides were found to contain 36% of behenic acid alongwith oleic (15.6%), linoleic (37.7%) and other fatty acids (10.5%) witha slip melting point of 40° C.

Example 5

[0034] Sunflower oil (2.5 g) and 1.22 g of ethyl behenate were taken ina screw-capped test tube and the mixture was homogenized by agitationfor 15 min. at 60° C. About 75 mg (2% by wt. of the substrate) ofLipozyme 1M 20 was added followed by agitation of the mixture for 3 hrat 60° C. After the reaction, the enzyme was filtered out and the enzymewas washed with hexane for its re-use. The product was purified bysilicic acid (60-120 mesh) column chromatography. The ethyl esters wereeluted by using hexane:ethyl acetate (99:1 v/v) followed bytriglycerides using a solvent mixture of hexane:ethyl acetate (95:5,v/v). The triglycerides were found to contain 34.7% of behenic acidalong with oleic (17.1%), linoleic (35.2%) and other fatty acids (13.0%)with a slip melting point of 37° C.

Example 6

[0035] Sunflower oil (2.5 g) and 1.15 g of methyl behenate were taken ina screw-capped test tube and the mixture was homogenized by agitationfor 15 min. at 60° C. About 73 mg (2% by wt. of the substrate) ofLipozyme 1M 20 was added followed by agitation of the mixture for 3 hrat 60° C. After the reaction, the enzyme was filtered out and the enzymewas washed with hexane for its re-use. The product was purified bysilicic acid (60-120 mesh) column chromatography. The methyl esters wereeluted by using hexane:ethyl acetate (99:1, v/v) followed bytriglycerides using a solvent mixture of hexane:ethyl acetate (95:5,v/v). The triglycerides were found to contain 16.5% of behenic acidalong with oleic (23%), linoleic (47.9%) and other fatty acids (12.6%)with a slip melting point of 20° C.

Example 7

[0036] Sunflower oil (2.5 g) and 1.28 g of isopropyl behenate were takenin a screw-capped test tube and the mixture was homogenized by agitationfor 15 min. at 60° C. About 76 mg (2% by wt. of the substrate) ofLipozyme 1M 20 was added followed by agitation of the mixture for 3 hrat 60° C. After the reaction, the enzyme was filtered out and the enzymewas washed with hexane for its re-use. The product was purified bysilicic acid (60-120 mesh) column chromatography. The isopropyl esterswere eluted by using hexane:ethyl acetate (99:1, v/v) followed bytriglycerides using a solvent mixture of hexane:ethyl acerate (95:5,v/v). The triglycerides were found to contain 5.1% of behenic acid alongwith oleic (26.5%), linoleic (60.0%) and other fatty acids (8.4%) with aslip melting point of <15° C.

Example 8

[0037] Sunflower oil (2.5 g) and 1.31 g of n-butyl behenate were takenin a screw-capped test tube and the mixture was homogenized by agitationfor 15 min. at 60° C. About 76 mg (2% by wt. of the substrate) ofLipozyme 1M 20 was added followed by agitation of the mixture for 3 hrat 60° C. After the reaction, the enzyme was filtered out and the enzymewas washed with hexane for its re-use. The product was purified bysilicic acid (60-120 mesh) column chromatography. The n-butyl esterswere eluted by using hexane:ethyl acetate (99:1, v/v) followed bytriglycerides using a solvent mixture of hexane:ethyl acetate (95:5,v/v). The triglycerides were found to contain 14.2% of behenic acidalong with oleic (23%), linoleic (52.3%) and other fatty acids (10.5%)with a slip melting point of 18° C.

Example 9

[0038] Sunflower oil (2.5 g) and 1.22 g of ethyl behenate were taken ina screw-capped test tube and the mixture was homogenized by agitationfor 15 min. at 70° C. About 75 mg (2% by wt. of the substrate) ofLipozyme 1M 20 was added followed by agitation of the mixture for 3 hrat 70° C. After the reaction, the enzyme was filtered out and the enzymewas washed with hexane for its re-use. The product was purified bysilicic acid (60-120 mesh) column chromatography. The ethyl esters wereeluted by using hexane:ethyl acetate (99:1, v/v) followed bytriglycerides using a solvent mixture of hexane:ethyl acetate (95:5,v/v). The triglycerides were found to contain 19.3% of behenic acidalong with oleic (20.5%), linoleic (48.5%) and other fatty acids (11.7%)with a slip melting point of 22° C.

Example 10

[0039] Sunflower oil (2.5 g) and 1.22 g of ethyl behenate were taken ina screw-capped test tube and the mixture was homogenized by agitationfor 15 min. at 70° C. About 75 mg (2% by wt. of the substrate) ofLipozyrne 1M 20 was added followed by agitation of the mixture for 3 hrat 55° C. After the reaction, the enzyme was filtered out and the enzymewas washed with hexane for its re-use. The product was purified bysilicic acid (60-120 mesh) column chromatography. The ethyl esters wereeluted by using hexane:ethyl acetate (99:1, v/v) followed bytriglycerides using a solvent mixture of hexane:ethyl acetate (95:5,v/v). The triglycerides were found to contain 19.3% of behenic acidalong with oleic (21.5%), lirioleic (51.9%) and other fatty acids(11.7%) with a slip melting point of 20° C.

Example 11

[0040] Safflower oil (2.5 g) and 1.22 g of ethyl behenate were taken ina screw-capped test tube and the mixture was homogenized by agitationfor 15 min. at 60° C. About 75 mg (2% by wt. of the substrate) ofLipozyme 1M 20 was added followed by agitation of the mixture for 3 hrsat 60° C. After the reaction, the enzyme was filtered out and the enzymewas washed with hexane for its re-use. The product was purified bysilicic acid (60-120 mesh) coluitun chromatography. The ethyl esterswere eluted by using hexane:ethyl acetate (99:1, v/v) followed bytriglycerides using a solvent mixture of hexane:ethyl acetate (95:5,v/v). The triglycerides were found to contain 24.7% of behenic acidalong with oleic (12.4%), linoleic (51.5%) and other fatty acids (11.4%)with a slip melting point of 28° C.

Example 12

[0041] Groundnut oil (2.5 g) and 1.22 g of ethyl behenate were taken ina screw-capped test tube and the mixture was homogenized by agitationfor 15 min. at 60° C. About 75 mg (2% by wt. of the substrate) ofLipozyme 1M 20 was added followed by agitation of the mixture for 3 hrat 60° C. After the reaction, the enzyme was filtered out and the enzymewas washed with hexane for its re-use. The product was purified bysilicic acid (60-120 mesh) column chromatography. The ethyl esters wereeluted by using hexane:ethyl acetate (99:1, v/v) followed bytriglycerides using a solvent mixture of hexane:ethyl acetate (95:5,v/v). The triglycerides were found to contain 22.0% of behenic acidalong with oleic (39.5%), linoleic (19.7%) and other fatty acids (18.8%)with a slip melting point of 25° C.

[0042] One embodiment of the invention relates to providing a lowcalorie fat composition comprising: Fatty Acid Wt % Palmitic 4.9 Stearic4.8 Oleic 17.1 Linoleic 35.2 Arachidic 1.7 Behenic 34.7 Lignoceric 1.6

[0043] Preferably, the fat composition comprising the following solidFat index: Temperature (° C.) SFI 15 11.5 20 9.9 25 9.4 30 8.2 35 3.4 400.0

[0044] Typically, the physical properties of the composition are:Physical Properties Saponification value 179.8 Iodine value 75.9 Acidvalue <0.1 M.Pt. (° C.) 37.0

[0045] In another embodiment, the fatty acid composition comprising thefollowing triglycerides: Triglyceride composition (by HPLC) ECN*Expected TAG** Wt % C42 LLL 12.1 C44 LLO 10.8 C44 LLP 2.8 C46 SLL; POL 3C46 LOO 2.4 C48 SOL; OOO 2.6 C50 BLL 32.3 C52 BLO 12.2 C52 BLP 2.4 C54BOO 1.9 C54 BOP 2.1 C56 BOS 1.7 C58 BLB 11.4 C60 BOB 2.1

[0046] The structured lipid synthesized enzymatically under thecondition of 2% of lipozyme (w/w of total substrate) at 60° C. for 3 hwas considered in this chapter for estimation of its calorific value (SL3). A 1 kg scale preparation of this structured lipid was carried outfor this purpose.

[0047] Almost all the products of normal lipid digestion and absorptionappear in the lymph. Shortly after a fatty meal, there is a greatincrease in the level of lipids in the lymph. This elevated lipid burdenof the lymph and hence the blood has been measured in several ways: astotal lipid, as triacylglycerol constituting 90% of the lymph lipid andalso as light-scattering particles (chylomicrons). Based on thisprinciple, single oral dose experiment of the proposed structured lipidhas been conducted on overnight fasting male Wistar rats. The absorptionbehavior of the structure lipid (SL 3) has been compared with that ofsunflower oil (control) by estimating and comparing the plasmatriacylglycerol level over a certain period of Lime (zero h, 1 h, 2 hand 3 h after oral dosing). In both the control and the experimentalgroup, the peak value of plasma triacyiglycerol was found to be 2 hafter loading of the fat. The level of plasma triacylglycerol was foundto be significantly higher in the control group than the experimentalgroup. The plasma as well as fecal total lipid fatty acid compositionwas also analyzed by gas-liquid chromatography to find the absorptionpattern of individual fatty acids. No trace of behenic acid was observedin the plasma total lipid of the experimental group and total excretionof behenic acid was found in the experimental group. Amount of excretedlipid in the fecal material of the experimental group was found to besignificantly higher than that of the control group. The encouragingresult of this experiment has helped in designing the nutritionalstudies meant for calorific estimation.

[0048] A modified growth method was adopted for estimating the calorificvalue of the studied structured lipid taking sunflower oil as caloriccontrol. The experiment was conducted on weanling NIN/Wistar male ratsof initial body weights 65-70 g and age 28 days. AIN-93 G dietformulated for growth was considered as basal diet of all the testgroups including the ad libitum group. Each test group along with adlibitum group contains seven numbers of rats. Basal feed consumption ofthe test groups is restricted to 50% of the feed consumed by the adlibitum group. Thus, all test groups receive equal amount of basal dietdaily. These diets were modified by adding 5, 10 and 15% of sunfloweroil for the control test groups, and 5 and 10% of studied structuredlipid for the experimental test groups. Daily adjustment of the feedquantities helps in maintaining rapid growth. The experiment wasconducted over 21 days. Body weights were recorded on 0, 3, 6, 9, 14, 17and 21 days. To find the linear response of the body weight gain of thecontrol animal against calorie intake, regression analysis wasperformed. Regression analysis was also performed to find the linearresponse of the body weight gain over 21 days of the conductedexperiment. The calorific value of the structured lipid was determinedby comparing the 21^(st) day mean growth of the rats fed with dietscontaining structured lipid to the 21^(st) day mean growth of the ratsfed with diets containing varying levels of sunflower oil. After thecumulative weight gain was recorded for 21 days, regression analysis wasperformed with added calories from sunflower oil regressed against21^(st) day body weight gains for the sunflower oil supplemented group.The slope and intercept calculated from the standard curve regressionwere used in the following formula, to estimate the calorific value ofthe structured lipid:${KCal}_{x} = \frac{{BWG}_{x} - {INT}}{{SLP} \times K_{x}}$

[0049] where KCal_(x) is the estimated Kilocalories per gram of unknownoil i.e. the structured lipid in this assay. BWG_(x) is the mean bodyweight gain on 21^(st) day for rats fed diets supplemented with 5% ofstructured lipid, INT is the intercept, SLP is the slope and K_(x) isthe amount of structured lipid added to the diet (grams per 100 g of thediet). An estimated value of 5.36 Kcal/g of the studied structured lipidwas found against 9 Kcal/g provided by a natural oil or fat. After 21days, blood as well as fecal samples were collected from all the groupsfor total lipid fatty acid composition analysis. The absorption patternof individual fatty acids in this long term feeding experiment wereidentical to those observed in the single oral dose experiment. Two suchgrowth studies were conducted.

[0050] Another growth study on ad libitum diet was also conducted on 21days old weanling NIN/Wistar male rats having their initial body weightsin the range of 39-40 g to find the metabolic behavior or the absorptionpattern of the structured lipid. Twenty rats were divided into twogroups. The AIN-93 G diet was taken as basal diet supplemented with 10%of sunflower oil for the control group and 10% of structured lipid forthe experimental group. The experiment was conducted over 28 days andbody weights were recorded on 0, 3, 5, 7, 14, 17, 19 and 21 days. Thegrowth patterns of both the groups were found to be identical. Mean foodintake of both the groups was also found to be identical in this adlibitum feeding experiment (9.8 g/day/rat for the control group and 9.9g/day/rat for the experimental group. After 28 days of the experiment,blood sample as well as fecal samples were collected from both thegroups for the total lipid fatty acid composition analysis. Nosignificant difference in the level of major fatty acids were foundbetween the two groups, though behenic acid was the predominant fattyacid excreted in the fecal material of the experimental group. All theseresults show that the absorption pattern of this semisynthetic studiedstructured lipid is very similar to natural oil or fat.

[0051] The following are the main observations:

[0052] i. Incorporation of behenic acid into vegetable oil was achievedboth by chemical interesterification and enzymatic transesterification.

[0053] ii. Structured lipid (SL 3), obtained by enzymatictransesterification between sunflower oil & ethyl behenate, provide 5.36Kcal/g against 9 Kcal provided by native oil or fat.

[0054] iii. Lower caloric availability of SL 3 is because of the totalexcretion of behenic acid in the feces.

[0055] iv. Identical growth pattern and mean food intake of control andexperimental group as found in growth study on ad libitum diet, confirmsthat the taste, palatability and absorption pattern of SL 3 is verysimilar to native oil or fat

[0056] v. The proposed structured lipid contain essential fatty acidsand natural antioxidants and hence nutritionally better equipped thanthe reduced calorie structured lipids, available in the market. TABLE 1Amount of total lipid excreted in the fecal material in the first growthexperimental on restricted diet Type of fecal fat Test group Mean ± SE(n = 3)* [mg/day] Glycerol lipids Control 1.343 ± 0.54 Experimental16.59 ± 3.27^(a) Fatty Acid salts Control  7.82 ± 1.14 Experimental52.72 ± 4.46^(b)

[0057] Growth study on restricted diet with 5% and 10% of structuredlipid: Table 1A graphically illustrates the calories delivered by allthe test groups. The two basic assumptions of this assay, that theweight gain of the rats is linearly related to kilo calories added tothe diet and the weight gain is linear over 14 days of the experiment,are confirmed from Table 8. In Table 2, the mean cumulated weight gainof the five test groups on restricted diet were recorded for each of thetime intervals when body weights were recorded over 21 days of theconducted experiment. Unlike Finley's assay (116), we have prolonged ourexperiment over 21 days. The slopes, intercepts and regressioncoefficients (r²) are also reported in Table 2. When mean body weightgain was regressed against 21 days of assay for all the groups,regression coefficients were found to be greater than 0.987. Thisfulfills the first basic assumption of this assay. Similarly, mean bodyweight gain of the control groups fed varying levels of sunflower oilwas also regressed against kilo calories added to the diet on each daywhen the body weights were recorded over 21 days of the conductedexperiment. Linearity has been observed from the 9^(th) day of the studyregression analysis saisfying the other basic assumption of this assay.When 21^(st) day mean body weight gain of all the control groups wereregressed against kilo calories added to the diet, a regressioncoefficient of 0.993, intercept of 21.13 and slope of 0.2111 wereobtained. TABLE 2 Mean body weight gains for various days and alsoagainst Keal added to the diet. Days of study regression analysis (± SE)Groups 0-3 0-6 0-9 0-14 0-17 0-21 Intercept^(a) Slope^(a,b) ^(a)r^(z) 5%SO  −1.4 ± 0.085  −0.78 ± 0.46  5.87 ± 0.67  9.75 ± 1.75   15 ± 1.75 21.5 ± 1.07 −7.07 1.35 0.987 (353 Kcal) 10% SO −1.187 ± 0.37    2.12 ±0.62  6.06 ± 0.79 11.87 ± 0.95 19.87 ± 1.18 25.68 ± 1.346 −7.29 1.590.994 (378 Kcal) 15% SO    0.5 ± 0.834    4.68 ± 1.01 10.75 ± 1.15716.62 ± 1.291 25.62 ± 1.451 32.05 ± 1.1498 −6.02 1.859 0.997 (403 Kcal)5% SL3 −2.629 ± 0.824 −0.375 ± 0.632  3.81 ± 0.875  6.75 ± 0.963 11.94 ±0.997 15.47 ± 1.801 −6.28 1.068 0.997 Intercept^(c) — — 5.12 9.312 14.821.13 Slope^(c) — — 0.098 0.1374  0.2124  0.2111 ^(c)r² — — 0.780 0.976 0.998  0.993

[0058] Estimation of calorific value of the structured lipid: As statedearlier, calorific availability of the structured lipid was determinedby comparing the 21^(st) day growth of the rats fed structured lipid tothe 21^(st) day growth of rats fed varying level of sunflower oil. Afterthe cumulative weight gain was determined for 21 days, regressionanalyses were performed with added calories from sunflower oil regressedagainst 21^(st) day body weight gains for the sunflower oil supplementedgroups. The slope and intercept calculated from the standard curveregression were used in the following formula to estimate the caloricvalue of the structured lipid. According to the formula,${KCal}_{x} = \frac{{BWG}_{x} - {INT}}{{SLP} \times K_{x}}$

[0059] where Kcal_(x) is the estimated kilocalories per gram of theunknown oil i.e. the structured lipid in this assay, BWG_(x) is the21^(st) day mean body weight gain for rats fed diets supplemented with5% of structured lipid, INT is the intercept from the standard curve,SLP is the slope of the standard curve and K_(x) is the amount ofstructured lipid added to the diet (grams per 100 g of the diet).Substituting all these values in the above equation, the estimatedkilocalorie per gram of the structured lipid is found to be,${{KCal}/g} = {\frac{15.47 - 21.13}{0.2111 \times 5} = {5.36\quad {{KCal}/g}}}$

[0060] Five rats were chosen randomly from each group for the fatty acidcomposition analysis of plasma total lipid contains the fatty acidcomposition in nanomoles per ml of plasma after 21 days of the conductedexperiment. The fatty acid composition in the experimental groupreflects dietary fatty acid intake, except that no trace of behenic acidwas observed. For example, there is significantly higher level of C18.2,C18.3, C20.0 and C20.3 in the plasma lipid of 5% control group than the5% experimental group, which is quite obvious. In 10% experiment, nosuch difference in the level of major fatty acid was observed betweenthe two groups, control and experimental. This indicates that at 10%added fat level, the required levels of all major fatty acids were metin the experimental group. Even in 10% experimental group, no behenicacid was observed.

[0061] Three rats were chosen randomly from all the groups of 5% and 10%experiment for the fatty acid composition analysis of the fecal lipid.Extraction and analysis was performed separately as neutral lipid andalso as soap.

[0062] A comparative presentation of the fatty acid composition of theexcreted lipids, both as neutral lipid and as soap, are given betweenthe control and experimental group in both 5% and 10% experiment. Dataindicates that there was a significantly higher amount of excretion ofall major fatty acids in the experimental group compared to the controlgroup. An absolute excretion of behenic acid as neutral lipid and mostlyas soap was also observed in the experimental group. No such excretionof behenic acid was found in the control group, which is a clearevidence of excreted behenic acid being a direct consequence of dietaryintake. A comparative picture of the amount of excreted lipid betweenthe control and experimental group in both the 5% and 10% experiment areshown below. Data depicts that there was a significantly higher amountof excretion of fecal lipid in the experimental group than the controlgroup (p<0.001). TABLE 2A A comparative picture of the amount ofexcreted lipid (mg/day) in fecal material Mean ± Type of experiment Typeof fecal lipid Groups SE (n = 3)*  5% Experiment Neutral Control  0.866± 0.2 Experimental  17.83 ± 14^(a) Control  3.62 ± 0.4 Soap Experimental 76.82 ± 7.6^(a) Control  1.93 ± 0.19 10% Experiment NeutralExperimental  7.38 ± 0.4^(a) Control  9.05 ± 0.8 Soap Experimental122.01 ± 7.9^(a)

[0063] Growth study on ad libitum diet supplemented with 10% structurallipid: No difference was attributed between the two groups in theirgrowth pattern over 21 days of the conducted experiment as is observedin Table 2B. This clearly depicts the similarity in the absorptionbehavior of structured lipid with the sunflower oil. Feed consumption ofboth the groups was recorded daily and here also no difference wasobserved between the two groups in their mean food intake. Mean foodintake of the control group is 9.8 g/day/rat and that of experimentalgroup is 9.9 g/day/rat.

[0064] This observation states that texture, flavor, and palatability ofthe structured lipid are very similar to natural oil.

[0065] All the rats from both the groups were chosen for the fatty acidcomposition analysis of plasma total lipid. In Table 3, the mean fattyacid composition of plasma total lipid in the control and theexperimental group are given in wt %. There was no difference in thelevel of all fatty acids in the plasma between the two groups. Thisfinding once again confirms the role played by the metabolic system inmaintaining the required level major fatty acids in the plasma. Threerats were chosen randomly from both the groups for their fecal lipidfatty acid composition analysis. Table 3 contains mean fatty acidcomposition of fecal lipid, neutral as well as soap, in both the controlgroup and the experimental group. TABLE 3 Analysis of fatty acidcomposition of plasma total lipid (in wt %) of the ad libitumexperiment; *Mean ± SE; n, number of rats. Fatty Acids Fatty Acid (wt %)Control* (n = 7) Experimental* (n = 7) 14:0 1.1 ± 0.1  1.2 ± 0.03 16:022.8 ± 0.4  21.7 ± 0.5  16.1 1.8 ± 0.4 2.7 ± 0.3 18:0 12.7 ± 0.6  12.3 ±0.6  18:1 14.1 ± 0.3  12.9 ± 0.4  18:2 19.5 ± 0.9  19.5 ± 0.3  20:0 0.5± 0.1 0.7 ± 0.1 18:3  0.2 ± 0.07 0.3 ± 0.1 20:3 1.6 ± 0.4   2 ± 0.2 20:423.2 ± 0.9  24.7 ± 0.4  22:0 — — 24:0 1.4 ± 0.2 0.8 ± 0.1 24:1 1.1 ± 0.11.2 ± 0.1

[0066] TABLE 4 Analysis of fatty acid composition of fecal total lipid(in wt. %) in the ad libitum experiment (10% added fat) Fatty Acid (wt%) Fatty Neutral Lipid (n = 3)* Soap (n = 3)* Acids Control ExperimentControl Experiment 14:0 1.3 ± 0.3  0.5 ± 0.03 1.2 ± 0.2  0.1 ± 0.03 16:0 21 ± 0.5   11 ± 0.6^(a) 26.1 ± 1.3   7.2 ± 0.6^(a) 18:0  26 ± 2.5 10.4± 0.03^(b) 30.2 ± 2.1   9.3 ± 0.5^(a) 18:1  22 ± 0.3  9.8 ± 1.8^(b) 16.5± 1.9    1 ± 0.6^(a) 18:2 10.6 ± 3.2   3.8 ± 1.2 6.9 ± 0.2  0.3 ±0.3^(b) 20:0 2.5 ± 0.2  3.9 ± 0.1 2.8 ± 0.3  4.1 ± 0.1^(b) 18:3 2.5 ±0.2  1.2 ± 0.3 1.4 ± 0.8  0.1 ± 0.1^(c) 20:3 7.6 ± 1   — 9.5 ± 0.8 —22:0 — 56.8 ± 3.6 — 75.6 ± 0.2 24:0 6.4 ± 0.2  2.6 ± 0.1^(b) 5.4 ± 0.3 2.3 ± 0.3^(b)

[0067] According to Table 4, there were significant increases in theamount of excretion of palmitic and stearic acid in the control groupthan the experimental group, both as neutral lipid and soap. There wasalso an absolute excretion of behenic acid in the experimental group andthe major excretion was as soap. There existed a significantly higheramount of excretion of fecal lipid as neutral lipid (p<0.01) and as soap(p<0.001) in the experimental group than the control group, as isobserved in Table 5. TABLE 5 Amount of excreted fecal lipid in the 10%SO (control group) and 10% SL3 (experimental) of the ad libitumexperiment Amount of excreted lipid Types of fecal lipid Types of testgroup (mg/day), n = 3* Neutral Control  2.9 ± 0.75 Experimental 22.97 ±4.36^(a) Soap Control  5.8 ± 2.1 Experimental 134.7 ± 9.7^(b)

ADVANTAGES OF THE PRESENT INVENTION

[0068] 1. The starting materials for the preparation of low caloriestructured fat are the natural edible oils preferably vegetable oilsources like mustard, sunflower, safflower and groundnut (peanut) oils.

[0069] 2. The process does nor involve any costly chemicals.

[0070] 3. The reduced-calorie plastic fat prepared in the presentinvention has all the physical, chemical and biological attributes of afat, consists only of triacylglycerols but deliver less calories anddoes not contain unnatural components such as very low molecular weightfatty acids.

[0071] 4. The reduced-calorie fat contains essential fatty acids andnatural antioxidants which are not present in the similar productsreported earlier.

[0072] 5. The reduced-calorie plastic fat does not contain any transfatty acids which are usually present in the similar type of plasticfats prepared by partial hydrogenation.

What is claimed is:
 1. A reduced calorie fat composition, whichcomprises: Fatty Acid wt % Palmitic about 4.9 Stearic about 4.8 Oleicabout 17.1 Linoleic about 35.2 Arachidic about 1.7 Behenic about 34.7Lignoceric about 1.6


2. The reduced calorie fat composition of claim 1, comprising thefollowing solid fat index: Temperature SFI 15° C. 11.5 20° C. 9.9 25° C.9.4 30° C. 8.2 35° C. 3.4 40° C. 0


3. The reduced calorie fat composition of claim 1, comprising thefollowing physical properties: Saponification Value 179.8 Iodine Value75.9 Acid Value <0.1 Melting Point about 37° C.


4. The reduced calorie fat composition of claim 1, comprising thefollowing triglyceride residues, as analyzed by HPLC: Effective CarbonNumber Expected TAG(s) wt % C 42 LLL 12.1 C 44 LLO 10.8 C 44 LLP 2.8 C46 SLL; POL 3 C 46 LOO 2.4 C 48 SOL; OOO 2.6 C 50 BLL 32.3 C 52 BLO 12.2C 52 BLP 2.4 C 54 BOO 1.9 C 54 BOP 2.1 C 56 BOS 1.7 C 58 BLB 11.4 C 60BOB 2.1,

where B represents a behenic acid residue, P represents a palmitic acidresidue, L represents a linoleic acid residue, O represents an oleicacid residue, and S represents a stearic acid residue.
 5. A reducedcalorie fat composition, which comprises palmitic acid, stearic acid,linoleic acid, arachidic acid, behenic acid, lignoceric acid, andresidues thereof.
 6. The reduced calorie fat composition of claim 5,which consists essentially of palmitic acid, stearic acid, linoleicacid, arachidic acid, behenic acid, lignoceric acid, and residuesthereof.
 7. The reduced calorie fat composition of claim 5, wherein thebehenic acid and its residue are present in an amount from about 29.8%to about 52.5% by weight of the composition.
 8. The reduced calorie fatcomposition of claim 5, wherein the behenic acid and its residue arepresent in an amount from about 5% to about 41% by weight of thecomposition.
 9. The reduced calorie fat composition of claim 5, whereinthe linoleic acid and its residue are present in an amount from about24.8% to about 44.6% by weight of the composition.
 10. The reducedcalorie fat composition of claim 5, wherein the linoleic acid and itsresidue are present in an amount from about 33% to about 60% by weightof the composition.
 11. A reduced calorie fat that is made by a processcomprising: esterifying one or more edible oils with 1,3-dibehenin inthe presence of a thermostable lipase enzyme at a temperature in therange of about 25° C. to 150° C. for at least about 0.5 hours; andrecovering the reduced calorie fat, wherein the thermostable lipaseenzyme is LIPOZYME.